퉁퉁마디로부터 색소체 외막 단백질 유전자의 분리 및 발현분석

네티 엘마와티,차준영,양영실,정민희,신동진,이병현,이곤호,손대영,Ermawati Netty,Cha, Joon-Yung,Liang, Yingshi,Jung, Min-Hee,Shin, Dongjin,Lee, Byung-Hyun,Lee, Kon-Ho,Son, Daeyoung 한국식물생명공학회 2004 식물생명공학회지 Vol.31 No.4

Differential display 방법으로 NaCl에 의하여 발현이 증가되는 cDNA들을 분리하였으며 그중 하나가 식물유래의 outer envelope membrane protein과 높은 유사성을 보였으므로 이를 ShOEP로 명명하였다. ShOEP는 1293 bp 길이에 359개의 아미노산으로 구성된 open reading frame을 포함하고 있으며, 이로부터 추정되는 분자량은 8.9 kDa이었다. ShOEP 단백질은 애기장대의 OEP와는 40.6%, 시금치와는 38%의 유사성을 나타내었다. Northern 분석결과, ShOEP 유전자는 NaCl의 농도가 증가함에 따라 발현량이 급격히 증가하는 것으로 나타났다. 염생식물인 퉁퉁마디의 OEP는 PEG에 의하여 발현이 증가하는 반면 비염생식물인 애기장대의 OEP는 큰 차이를 보이지 않았다. 효모 complementation 실험결과 ShOEP는 NaCl에 특이적이었으며 식물의 염분스트레스 내성 기작에 직접적으로 관여하고 있음을 알 수 있었다. Complementary DNA encoding chloroplast outer envelope membrane protein (OEP) from the halophyte Salicornia herbacea has been cloned and sequenced. The full length cDNA is 596 bp and encodes a polypeptide of 91 amino acid residues with a molecular mass of 8.9 kDa. The expression level of ShOEP increased by salt, drought and ABA treatments. ShOEP expression was largely induced in roots and shoots by high salts. The biological function of ShOEP was examined by yeast complementation. ShOEP can suppress Na$^{+}$ sensitivity of yeast mutant (cnb$\Delta$) in the presence of salt. These results suggest that ShOEP is a salt inducible gene and may have functions in the regulation of plant salt stress.ant salt stress.

퉁퉁마디로부터 색소체 외막 단백질 유전자의 분리 및 발현분석

네티 엘마와티,차준영,양영실,정민희,신동진,이병현,이곤호,손대영 Plant molecular biology and biotechnology research 2004 Plant molecular biology and biotechnology research Vol.2004 No.-

Complementary DNA encoding chloroplast outer envelope membrane protein (OEP) from the halophyte Salicornia herbacea has been cloned and sequenced. The full length cDNA is 596 bp and encodes a polypeptide of 91 amino acid residues with a molecular mass of 8.9 kDa. The expression level of ShOEP increased by salt, drought and ABA treatments. ShOEP expression was largely induced in roots and shoots by high salts. The biological function of ShOEP was examined by yeast complementation. ShOEP can suppress Na+ sensitivity of yeast mutant (cnbΔ) in the presence of salt. These results suggest that ShOEP is a salt inducible gene and may have functions in the regulation of plant salt stress.

퉁퉁마디로부터 염에 의하여 유도되는 Aldolase 유전자의 분리 및 발현분석

차준영,네티 엘마와티,김순길,이증주,임채오,정우식,이곤호,손대영 Plant molecular biology and biotechnology research 2003 Plant molecular biology and biotechnology research Vol.2003 No.-

Soil salinity is one of the most serious abiotic stresses limiting the productivity of agricultural crops. To cope with salt stress, plants respond with physiological, developmental and biochemical changes, including the synthesis of a number of proteins and the induction of gene expression. Salicornia herbacea is a halophytic plant that grows in salt marches and on muddy seashores. In order to understand the biochemical and molecular mechanisms of salt tolerance in S. herbacea, we isolated several genes that involved in the salt tolerance by mRNA differential display.Here we report the cloning of a cDNA encoding fructose-1, 6-bisphosphate aldolase, named ShADL, which is 1293 bp long and contains an open reading frame consisted of 359 amino acids with calculated molecular mass of 39 kDa. ShADL protein showed 86% identity with Arabidopsis and 78% with aldolase of common ice plant. Northern blot analysis revealed that the transcript of ShADL gene was increased dramatically depending on the NaCI concentrations.

단백체를 이용한 애기장대 Cytokinin 유도 단백질의 분석

양영실,차준영,네티 엘마와티,정민희,배동원,이창원,손대영,Liang Ying-Shi,Cha Joon-Yung,Ermawati Netty,Jung Min-Hee,Bae Dong-Won,Lee Chang-Won,Son Dae-Young 한국식물생명공학회 2005 식물생명공학회지 Vol.32 No.4

Cytokinins are essential plant hormones that play crucial roles in various aspects of plant growth and development. To better understand the molecular mechanisms of cytokinin action, we identified cytokinin related proteins by a proteomic approach. Proteins extracted from control and trans-zeatin treated Arabidopsis seedlings were separated and analyzed by two dimensional gel analysis. Differentially expressed protein spots were identified with peptide mass fingerprinting based on matrix assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry and database searching, We obtained ten up-regulated and one down-regulated proteins upon t-zeatin treatment. The expression of the following proteins was induced; pollen allergen like protein, L-ascorbate peroxidase, tetrapyrrole methylase family protein, SGT1 protein homolog, disease resistance related protein, maternal embryogenesis control protein, paxneb related protein, gluthathione S-transferase and IAA amino acid hydrolase homolog. Cytokinin은 식물의 성장과 발달에 중요한 역할을 하는 필수 호르몬이다. Cytokinin의 작용 기작을 이해하기 위하여 단백체를 이용하여 cytokinin 관련 단백질들을 동정하였다. 대조구와 t-zeatin을 처리한 애기장대로부터 추출한 단백질을 이차원 전기영동하여 분석하였다. 발현양에 차이가 있는 단백질 spot들을 MALDI-TOF 단백질 질량분석기와 database 검색을 통하여 동정하였다. 그 결과 t-zeatin 처리에 의하여 발현이 증가하는 10개의 단백질과 감소하는 한 개의 단백질을 분리할 수 있었다. Cytokinin에 의하여 발현이 증가하는 단백질은 pollen allergen like protein, L-ascorbate peroxidase, tetrapyrrole methylase family protein, SGT1 protein homolog, disease resistance related protein, maternal embryogenesis control protein, paxneb related protein, gluthathione S-transferase, 그리고 IAA amino acid hydrolase homolog들로 밝혀졌다.

Characterization of Arabidopsis Histidine Kinase 3 and Proteomic Analysis of Its Mutant

양영실,차준영,네티 엘마와티,정민희,이곤호,손대영,Liang Ying-Shi,Cha Joon-Yung,Ermawati Netty,Jung Min-Hee,Lee Kon-Ho,Son Dae-Young Korean Society of Life Science 2006 생명과학회지 Vol.16 No.3

Histidine kinase는 식물의 신호전달기작에서 매우 중요한 역할을 한다. 본 연구에서는 애기장대 histidine kinase 3 (AHK3)의 식물체내에서의 기능을 조사하였으며 이 유전자의 결손 돌연변이체인 ahk3에 trans-zeatin (t-zeatin)을 처리하여 유전자와 단백질의 발현양상을 분석하였다. ahk3는 야생형 식물체에 비하여 캘러스 형성, 유모의 성장, 잎의 노화과정에서 t-zeatin에 대한 감수성이 줄어들었다. 프로테옴 분석 결과 eukaryotic translation initiation factor 5A-2, auxin binding glutathione S-transferase, NDPK1 등은 야생형의 애기장대에서는 t-zeatin에 의하여 발현이 증가하는 반면 ahk3에서는 증가하지 않는 것으로 나타났다. 또한 cytokinin처리에 의하여 발현이 증가하는 것으로 보고된 A-type response regulator들 중에서 ARR4와 ARR16의 발현양이 ahk3에서는 현저하게 감소하는 것으로 나타났다. 이러한 결과들은 AHK3가 cytonin신호전달기작에서 매우 중요한 역할을 하며, 프로테옴 분석에 의하여 동정된 단백질들과 ARR4, ARR16은 AHK3에 의해 매개되는 cytokinin 신호전달과정에서 중요한 역할을 할 것으로 생각된다. Histidine kinase plays important roles in signal transduction in plant. We characterized the function of Arabidopsis histidine kinase 3 (AHK3) and analyzed the expression patterns of genes and proteins in its mutant ahk3 by trans-zeatin (t-zeatin). The ahk3 exhibited decreased sensitivity to t-zeatin during callus formation, seedling growth, and leaf senescence. From proteomic analysis of ahk3, eukaryotic translation initiation factor 5A-2, auxin binding glutathione S-transferase, and NDPK1 were identified not to be induced by t-zeatin, when compared to the wild-type. In addition, the expression levels of ARR4 and ARR16 among A-type response regulators (ARRs) markedly decreased in ahk3 by t-zeatin treatment. These results suggest that AHK3 plays an important role in cytokinin signaling and the proteins identified from proteomic analysis and specific ARRs, ARR4 and ARR16 may be directly or indirectly associated in AHK3-mediated cytokinin signaling.

단백체를 이용한 애기장대 Cytokinin 유도 단백질의 분석

양영실,차준영,네티 엘마와티,정민희,배동원,이창원,손대영 Plant molecular biology and biotechnology research 2005 Plant molecular biology and biotechnology research Vol.2005 No.

Cytokinins are essential plant hormones that play crucial roles in various aspects of plant growth and development. To better understand the molecular mechanisms of cytokinin action, we identified cytokinin related proteins by a proteomic approach. Proteins extracted from control and trans-zeatin treated Arabidopsis seedlings were separated and analyzed by two dimensional gel analysis. Differentially expressed protein spots were identified with peptide mass fingerprinting based on matrix assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry and database searching. We obtained ten up-regulated and one down-regulated proteins upon t-zeatin treatment. The expression of the following proteins was induced; pollen allergen like protein, L-ascorbate peroxidase, tetrapyrrole methylase family protein, SGT1 protein homolog, disease resistance related protein, maternal embryogenesis control protein, paxneb related protein, gluthathione S-transferase and IAA amino acid hydrolase homolog.

Hsp90 샤페론 복합체내 식물 p23의 역할

차준영,무하마드 수디,양영실,정민희,네티 엘마와티 한국초지조사료학회 2005 한국초지조사료학회 학술대회논문집 Vol.2005 No.1